A solid-solution theory of anesthetic interaction with lipid membranes: temperature span of the main phase transition

Anesthetics (or any other small additives) depress the temperature of the main phase transition of phospholipid bilayers. Certain anesthetics widen the temperature span of the transition, whereas others do not. The widening in a first-order phase transition is intriguing. In this report, the effects of additive molecules on the temperature and its span were explained by the solid-solution theory. By assuming coexistence of the liquid-crystal and solid-gel phases of lipid membranes at phase transition, the phase boundary is determined from the distribution of anesthetic molecules between the liquid-crystal membrane versus water and between the solid-gel membrane versus water. The theory shows that when the lipid concentration is large or when the lipid solubility of the drug is large, the width of the transition temperature increases, and vice versa. Highly lipid-soluble molecules, such as long-chain alkanols and volatile anesthetics, increase the width of the transition temperature when the lipid:water ratio is large, whereas highly water-soluble molecules, such as methanol and ethanol, do not. The aqueous phase serves as the reservoir for anesthetics. Depletion of the additive molecules from the aqueous phase is the cause of the widening. When the reservoir capacity is large, the temperature width does not increase. The theory also predicts asymmetry of the specific heat profile at the transition.     

Effect of Silicate Grain Shape, Structure, and Location on the Biomass and Community Structure of Colonizing Marine Microbiota

Microbiota colonizing silica grains of the same size and water pore space, but with a different microtopography, showed differences in biomass and community structure after 8 weeks of exposure to running seawater. The absence of surface cracks and crevices resulted in a marked diminution of the total microbial biomass measured as lipid phosphate and total extractable palmitic acid. With increasing smoothness of the sand grain surface, examination of the community structure showed a marked decrease in procaryotes and algal microeucaryotes, with a relative increase in microeucaryotic grazers. A comparison of the colonizing sediment incubated in running seawater or at 32 m on the sea floor with a sediment core showed a decreased bacterial biomass with a different community structure and a decreased total microeucaryotic population of both grazers and algae. The quantitative differences in microbial biomass and community structure between the microcosms and the actual benthic population in the core were determined.     

Effect of a taurine treatment on the regression of existing atherosclerotic lesions in rabbits fed on a high-cholesterol diet

We studied whether taurine has any regressive effect on existing atherosclerotic lesions and lipid peroxidation in rabbits fed on a high-cholesterol (HC) diet. The cholesterol, triglyceride, malondialdehyde (MDA) and diene conjugate (DC) levels, as well as the aortic histopathological findings were examined in rabbits that had been fed on a cholesterol-containing diet for 8 months [0.5% cholesterol (w/w) for 3 months and subsequently 0.25% cholesterol (w/w) for 5 months], and then for a further 4 months on a normal diet with or without taurine treatment [1% (w/v) in the drinking water]. High levels of lipid and lipid peroxide induced by the HC diet were observed to decline in the plasma, liver and aorta of atherosclerotic rabbits, as well as a slight retardation in aortic atherosclerotic lesions during the regression period. Although no significant differences in the lipid and lipid peroxide levels in the plasma and aorta were found between the regressed groups with or without the taurine treatment, the extent of atherosclerotic lesions in the aorta was less in the taurine-treated regressed group than in the non-treated regressed group. However, the liver MDA and DC levels were lower in the regressed rabbits with the taurine treatment in the non-treated group. These results indicate that the taurine treatment may accelerate the regression of cholesterol-induced atherosclerotic lesions in rabbits without having any effect on the plasma and aorta lipid and lipid peroxide levels.     

Conformation of bombesin in buffer and in the presence of lysolecithin micelles: NMR, CD, and fluorescence studies

The conformation of the tetradecapeptide hormone bombesin has been studied in buffer and in the presence of lysolecithin micelles, using static and dynamic fluorescence, CD, and one- and two-dimensional nmr. The results obtained show that in buffer bombesin is present in an extended flexible chain, with no evidence for any ordered secondary structure. A marked change in the CD spectrum is observed changing from buffer to the lipid suspension. Concomitantly, the 1H-nmr spectrum of bombesin, in a D2O lipid dispersion, shows the persistence of resonances due to exchangeable protons and in similar conditions the fluorescence intensity increases. We think therefore that these results strongly support the hypothesis that bombesin interacts with the lipid phase, assuming ordered secondary structure. Finally, the marked dependence of tryptophan fluorescence quantum efficiency and order parameter from the hormone concentration in the presence of lysolecithin but not in buffer leads to the conclusion that bombesin can associate into the lipid matrix.     

Can physicochemical factors predict lipid content in phytoplankton?

1. We hypothesized that a large collection of reasonably standardized data for natural algal assemblages would reveal the influence of environmental factors on the fraction of recently produced photosynthate allocated to lipid.
2. Our analysis of photosynthate allocation in fresh- and saltwater systems showed that allocation of carbon to lipid was not well correlated with any one environmental factor.
3. Allocation to lipid increases with temperature up to 12 °C (lipid allocation = 11.7 + 1.70*temperature (°C), n = 48, r ² = 0.50, P < 0.001) but not above.
4. The relationship between temperature and lipid allocation appears to be caused by a unique convergence of nutrient limitation (nitrate or silicate) in the presence of facultative lipid-producing algae (diatoms or other chrysophytes) which occurs at or below 12 °C during stratification of the water column.     

Kinetics of charge transfer at the lipid bilayer-water interface on the nanosecond time scale.

Advances in instrumentation allow electrical measurements across the planar lipid bilayer to be made with nanosecond time resolution. The electron transfer reaction between photoexcited magnesium octaethylporphyrin in the lipid to a variety of ionically charged acceptors in the water is found to be purely dynamic over a wide range of concentrations of acceptors and up to the time constant of the apparatus, 4 ns. The saturation of the amplitude of the photovoltage with increasing concentration of acceptor is caused by the finite lifetime of the excited state, not by formation of a static pigment-acceptor complex. The reactions are an excellent probe of the lipid-water interface over an extended time scale. No appreciable barrier to reaction exists at this interface beyond the 5-ns time. That is, any water or choline group structure may be evanescent on this time scale. Electrostatic interactions indicate that the acceptor molecules penetrate to the level of the phosphocholine groups with differing orientations. It will be possible to extend the time scale into the picosecond range by decreasing the response time and by deconvolutions.     

Changes in the proximate body composition of the landlocked sea lamprey Petromyzon marinus (L.) during larval life and metamorphosis

Estimates were made of the growth rates and proximate body composition of larval and metamorphosing P. marinus (L.) collected at various times of the year from Shelter Valley Creek, Lake Ontario. Analysis of length-frequency data indicates that the average duration of larval life was 6 years, with metamorphosis occurring predominantly in the length range above 13 cm. Increases in length were almost entirely restricted to the warmest months and did not take place during the final year of larval life. Three categories were thus recognized for the proximate analysis: ammocoetes <13 cm, ammocoetes > 13 cm and metamorphosing individuals. In ammocoetes <13 cm, seasonal differences were observed in the regression coefficients in the logarithmic relationships between wet weight and length and between each of water, lipid and ash and the wet weight. No such difference was found for the regressions between protein and wet weight. For a fixed length (9 cm), the wet weight varied only slightly during the year, although a small peak was seen in May. When considered on the basis of fixed weight (1 g), the relative amount of lipid deposited was greatest in May/July, coincident with a high diatom density. The water content followed an inverse pattern to that of lipid, while the protein and ash contents showed little seasonal variation and exhibited values lower than those normally found in teleosts. Throughout the last year of larval life, the animal stored a greater proportion of lipid, presumably to facilitate the energy demands of metamorphosis during which this food store underwent a marked reduction.     

Seasonal changes in particulate and dissolved lipids in a eutrophic prairie lake

1. Critical periods of lipid energy transfer from phyto- to zooplankton were inferred by comparing seasonal patterns of particulate and dissolved lipid fractions in lake water with temporal changes in lipid energy reserves of the zooplankton in a hypereutrophic lake.
2. The midsummer phytoplankton community was dominated by the bloom-forming cyanobacterium Aphanizomenon flos-aquae. The collapse of the bloom was accompanied by a 2-week period of severe nitrogen deficiency after which there was a marked increase in the concentration of lipid energy reserves in the particulate (algal) fraction.
3. Areal lipid energy reserves of the dominant herbivorous zooplankton responded positively to changes in the tri-and diacylglycerol content of the particulate fraction of lake water in a species-specific manner.
4. Bacterial numbers also peaked in September concomitant with a large increase in free fatty acids in the dissolved lipid fraction probably produced by the decay of the
A. flos-aquae bloom.
5. The association between periods of nitrogen deficiency and increased energy reserve lipids in the particulate fraction supports observations made with laboratory algal cultures that periods of nutrient deficiency may intensify lipid synthesis in some algal species, thereby enhancing the rate of lipid energy transfer from phytoplankton to zooplankton.     

Membrane Lipids in the Aging Human Brain

The membrane lipid composition of human brain has been studied in 21 men and 18 women 60-97 years of age. This brain tissue series is unique, because it has been obtained only from individuals who lived a normal social life in their own homes and died suddenly and unexpectedly from arteriosclerotic heart disease or ruptured aortic aneurysms. They had no history of neurological or psychiatric disease. Macroscopic and microscopic examinations ruled out any signs of organic brain disorder. The percentage of solids diminished continuously during the whole period, but the marked individual differences suggested large variations in the hydration of the brain. The content of membrane lipids also diminished continuously up to 90 years of age, when a marked diminution in level of gangliosides and cerebrosides occurred, a result indicating a rapid reduction in amount of neuronal membranes and myelin. The clinical implications of the variation in brain hydration and the rapid loss of membrane lipids after 90 years of age are discussed.     

A Model Representing a Physiological Role of CO2 at the Cell Membrane

A model is presented suggesting the interaction of CO₂ and bicarbonate on lipids of the cell membrane. The interfacial tensions between water and oil (benzene) phases were measured using the stalagmometer and the sessile drop methods. Effects of electrolyte solutions and of CO₂ on molecular arrangement at the interface were calculated. Chloride solutions against oleic acid in benzene produced little decrease in interfacial tension from that measured for pure water against the oil phase. Presence or absence of CO₂ caused no change in interfacial tension of water or chloride solutions against the oil phase. Bicarbonate salts in the absence of CO₂ caused marked decreases in interfacial tension from that measured for water or chloride solutions. Concomitant with this decrease in interfacial tension were an increase in hydration of the interface and changes in molecular spacings of the lipid. This hydration may be considered as reflecting a more ionic-permeable cell membrane. The addition of CO₂ to the bicarbonates caused an increase in interfacial tension of the model, approaching that of the chlorides, with decreased hydration of the interface. Viewed as occurring at the cell membrane this would make the lipid more continuous and decrease the ease of ionic penetration. In this way the action of bicarbonates and CO₂ at the interface suggests an explanation of the action of CO₂ on the cell.     

Oxytocin gene deletion mice overconsume palatable sucrose solution but not palatable lipid emulsions

We previously reported that oxytocin knockout (OT KO) mice display markedly enhanced intake of sweet and nonsweet carbohydrate solutions compared with intake by wild-type (WT) mice of the same background strain. The present study was conducted to determine whether OT KO mice demonstrate enhanced intake of Intralipid, a palatable lipid emulsion. Male or female mice of both genotypes that were naive to the test solution were given continuous two-bottle access to Intralipid and water with food available ad libitum for 3 days. Throughout the experiment, mice of both genotypes showed a marked preference for Intralipid over water. On the 1st day, OT KO mice displayed twofold greater preference and consumed nearly twice as much Intralipid compared with WT cohorts. However, on subsequent days of exposure, Intralipid preference and intake did not differ between genotypes over a range of lipid concentrations presented in descending or ascending order. Daily and hourly measures of lipid vs. sucrose intake confirmed that OT KO mice consumed more sucrose solution, but not lipid emulsion, than WT mice. During ad libitum access to Intralipid, both genotypes consumed significantly more calories from the emulsion as concentration increased. Both genotypes maintained consistent total daily caloric intake (lipid plus chow) and compensated by decreasing chow intake over the course of the study. These findings, coupled with prior reports from our laboratory, support the view that OT signaling pathways participate in limiting intake of palatable carbohydrate-containing solutions, but do not appear to play a role in limiting intake of Intralipid.     

Effect of ionol on the stomach lesion in rats during immobilization stress

Intragastric administration of the antioxidant ionol (50 mg/kg/a day) for 7 days did not prevent the formation of ulcers or massive hemorrhages to the gastric mucosa in immobilized animals, increased the number of petechiae and lowered that of erosions. In this case the parietal pH increased in the stomach and dropped in the remaining divisions of the alimentary tract. The combined action of stress and ionol brought about marked hypocoagulation. Ionol exerted a protective action on the stress-induced activation of lipid peroxidation in the stomach but did not produce any such effect on the liver.     

Valinomycin-mediated ion transport through neutral lipid membranes: Influence of hydrocarbon chain length and temperature

Summary Stationary electrical conductance experiments together with nonstationary relaxation experiments allow a quantitative determination of rate constants describing carrier-mediated ion transport. Valinomycin-induced ion transport across neutral lipid membranes was studied. The dependence of the transport parameters on the chain length of the lipid molecules, on the kind of alkali ion, and on the temperature was determined. The relaxation time the current following a voltage jump shows a marked increase with decreasing temperature or with increasing chain length of the lipid molecules. This variation of is interpreted on the basis of a varying membrane fluidity. It is shown that under favorable circumstances the equilibrium constant of complex formation in the aqueous phase may be obtained from membrane experiments. Furthermore, the kinetics of exchange of valinomycin between membrane and water was studied. We found a marked influence of the totus surrounding the black film on the kinetics as well as on the total amount of valinomycin molecules in the membrane. The problem of location of the free carrier molecules inside the membrane is discussed.     

Age-related changes in the morphology and function of the zona glomerulosa of the rat adrenal cortex

The age-related changes in the morphology and function of rat adrenal zona glomerulosa (ZG) were investigated by coupled stereological and radioimmunological techniques. For this purpose 4-, 8-, 16- and 24-month-old rats were studied. Aging caused a notable lowering in the plasma aldosterone concentration and a marked decrease in both basal and ACTH- or angiotensin II (ANG-II)-stimulated secretion of collagenase-dispersed ZG cells. Plasma renin activity (PRA) underwent an age-dependent decrease, while the plasma level of ACTH displayed a significant rise. ZG and its parenchymal cells did not evidence any age-related morphologically demonstrable alteration in their growth, nor ZG cells showed any marked ultrastructural change, with the exception of a severe lipid-droplet repletion. This last finding is in keeping with the aging-induced decrease in the secretory activity of ZG cells, inasmuch as lipid droplets are the intra-cellular stores of cholesterol esters, the obligate precursors of steroid hormones in rat adrenals. ACTH and ANG-II are well known to be involved in the maintenance of the growth of rat ZG; thus, the combined impairment of ANG-II production (as evidenced by PRA lowering) and increase in ACTH secretion may maintain unchanged ZG growth during aging.     

Phosphate-limited oat. The plasma membrane and the tonoplast as major targets for phospholipid-to-glycolipid replacement and stimulation of phospholipases in the plasma membrane

We recently reported that cultivation of oat (Avena sativa L.) without phosphate resulted in plasma membrane phosphoglycerolipids being replaced to a large extent by digalactosyldiacylglycerol (DGDG) (Andersson, M. X., Stridh, M. H., Larsson, K. E., Liljenberg, C., and Sandelius, A. S. (2003) FEBS Lett. 537, 128-132). We report here that DGDG is not the only non-phosphorous-containing lipid that replaces phospholipids but that also the content of glucosylceramides and sterolglycosides increased in plasma membranes as a response to phosphate starvation. In addition, phosphate deficiency induced similar changes in lipid composition in the tonoplast. The phospholipid-to-glycolipid replacement apparently did not occur to any greater extent in endoplasmic reticulum, Golgi apparatus, or mitochondrial inner membranes. In contrast to the marked effects on lipid composition, the polypeptide patterns were largely similar between root plasma membranes from well-fertilized and phosphate-limited oat, although the latter condition induced at least four polypeptides, including a chaperone of the HSP80 or HSP90 family, a phosphate transporter, and a bacterial-type phosphoesterase. The latter polypeptide reacted with an antibody raised against a phosphate deficiency-induced phospholipase C from Arabidopsis thaliana (Nakamura, Y., Awai, K., Masuda, T., Yoshioka, Y., Takamiya, K., and Ohta, H. (2005) J. Biol. Chem. 280, 7469-7476). In plasma membranes from oat, however, a phospholipase D-type activity and a phosphatidic acid phosphatase were the dominant lipase activities induced by phosphate deficiency. Our results reflect a highly developed plasticity in the lipid composition of the plasma membrane and the tonoplast. In addition, phosphate deficiency-induced alterations in plasma membrane lipid composition may involve different sets of lipid-metabolizing enzymes in different plant tissues or species, at different stages of plant development and/or at different stages of stress adjustments.     

Polyunsaturated fatty acyl residues of galactolipids are involved in the control of bilayer/non-bilayer lipid transitions in higher plant chloroplasts

Total polar lipid extracts of chloroplasts isolated from broad beans (Vicia faba) tend to form non-bilayer structures when dispersed in dilute salt solutions. Monoglactosyldiacylglycerol is shown to play a dominant role in this process. The tendency of this lipid to form non-bilayer structures when dispersed alone in water was found to depend upon the degree of unsaturation of its associated fatty acyl chains. Highly unsaturated lipids (average number of double bonds per lipid molecule greater than about 5.0) form inverted hexagonal (Hex_II) structures in water at 20°C, whilst more saturated lipids (average number of double bonds per lipid molecule less than about 4.5) form lamellar sheets. Wide-angle X-ray diffraction and differential scanning calorimetry measurements indicate that these lamellae consist of gel-phase lipid that can adopt either of two structures depending on the thermal history of the sample. Freeze-fracture studies performed on total polar lipid extracts that have been hydrogenated using Adams' catalyst, and reconstituted extracts in which monogalactosyldiacylglycerol has been selectively hydrogenated, show that the degree of unsaturation of this lipid is a key factor in determining whether or not non-bilayer structures are formed in such extracts. Increasing the extent of saturation of the acyl residues of monogalactosyldiacylglycerol reduces the tendency to form non-bilayer structures. Similar effects are observed on lowering the temperature of the dispersions. Fluorescence polarisation measurements using 1,6-diphenyl-1,3,5-hexatriene indicate that the disappearance of non-bilayer structures is accompanied by a marked decrease in the fluidity of the lipid matrix. The possible significance of these observations is discussed in terms of the thermal adaptation and chilling sensitivity of plant membranes.     

The transmembrane protein bacterioopsin affects the polarity of the hydrophobic core of the host lipid bilayer.

Influence of the transmembrane protein bacterioopsin (the retinal-free form of bacteriorhodopsin) on the polarity of egg-phosphatidylcholine bilayers was studied by means of a steady-state and time-resolved fluorescence approach exploiting the solvatochromic properties of the 2-anthroyl fluorophore. Introduced in phosphatidylcholine molecules in the form of 8-(2-anthroyl)octanoic acid, this fluorophore probed the hydrocarbon core of the lipid bilayer. As previously shown (E. Pérochon et al., Biochemistry 31 (1992) 7672-7682), water molecules were detected in this region of the terminal part of the lipid acyl chains. Their number was considerably reduced upon addition of bacterioopsin to the lipids. This was assessed by a blue shift in the fluorescence emission spectra of the probe and a marked decrease in the fractional population of fluorophores interacting with water, to the benefit of those experiencing a hydrophobic environment. In agreement with current theories, this decrease in the hydration of the bilayer may be linked to an increase in the acyl chain order and a decrease in the lateral diffusion coefficient of lipids near the protein. The data obtained at high protein concentration accounts for a protein/lipid interface which is much less hydrated than the hydrophobic core of a protein-free lipid bilayer.     

A comparative cytological study between hepatocytes of insecticide-resistant and susceptible mosquitofish (Gambusia affinis).

The livers from a naturally occurring insecticide-resistant mosquitofish population were examined cytologically and compared to liver preparations from a susceptible population. There was a marked increase in cell size and lipid inclusions in the hepatocytes of resistant mosquitofish as compared to susceptible mosquitofish. The pronounced lipid inclusions in the liver preparations of resistant mosquitofish remained essentially unchanged in mosquitofish starved for up to 2 weeks. No observable differences were noted in any cellular organelles.     

Effects of cholesterol surface transfer on cholesterol and phosphatidylcholine synthesis in cultured rat arterial smooth muscle cells

The purpose of this study was to examine the effects of cholesterol surface transfer between lipid vesicles and rat arterial smooth muscle cells on endogenous synthesis of cholesterol and phosphatidylcholine. Lipid vesicles containing cholesterol and egg phosphatidylcholine in different proportions were used as the extracellular lipid source. The rate of cellular cholesterol and phosphatidylcholine synthesis was determined from the [14C]acetate incorporation into these lipid classes. [3H]Cholesterol in lipid vesicles, with a cholesterol/phospholipid (C/P) mole ratio of 1:1, was rapidly transferred into rat smooth muscle cells, with a half-time of about 3.6 hours in the absence of serum proteins. Incubation of cells for 5 hours with vesicles of a high C/P mole ratio (i.e. 1.5:1) at vesicle-cholesterol concentrations above 100 micrograms/ml resulted in a marked reduction of cellular cholesterol synthesis, whereas the rate of phosphatidylcholine synthesis was increased. Cells incubated with lipid vesicles of C/P 1:2 did not show any change in cellular cholesterol or phosphatidylcholine synthesis. Incubation of cells with egg phosphatidylcholine vesicles at concentrations above 300 micrograms/ml, on the other hand, stimulated endogenous synthesis of cholesterol without affecting cellular phosphatidylcholine synthesis. The main conclusion is that cholesterol surface transfer may influence cellular lipid metabolism in the absence of mediating serum lipoproteins in a model system with cultured cells and lipid vesicles.     

Changes in muscle, liver, and plasma free fatty acid levels in the pigeon on acute exposure to cold.

Adult pigeons were subjected to acute cold exposure (-25 degrees C; 30 min). Fully-plumed birds, showed a pronounced increase in the level of FFA in the blood, but not in the liver or muscle. Partially-defeathered (dorsum and pectoral regions) birds, likewise, showed a marked increase in plasma FFA level but failed to indicate any change in FFA levels in the liver or the muscle. It is concluded that even if the mobilized FFA may have supported calorigenic processes in the normothermic cold-exposed pigeon, lipid reserves are unlikely to have served as a significant source of energy for thermogenic functions in the hypothermic (defeathered) bird.